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Introduction Lack of reliable peak detection impedes automated analysis of large-scale gas chromatography-mass spectrometry (GCMS) metabolomics datasets. Performance and outcome of individual peak-picking algorithms can differ widely depending on both algorithmic approach and parameters, as well as data acquisition method. Therefore, comparing and contrasting between algorithms is difficult. Technological and methodological innovation We present part of the work published in [1] and implemented in our workflow for improved peak picking (WiPP), focusing on the use of machine learning-based classification to optimize and improve different steps of the common GC-MS metabolomics data processing workflow. Our approach evaluates the quality of detected peaks using a machine learning based classification scheme based on seven peak classes. The quality information returned by the classifier for each individual peak is merged with results from different peak detection algorithms to create one final high-quality peak set for immediate down-stream analysis. Results and impact We benchmarked our workflow to standard compound mixes and a complex biological dataset, demonstrating that peak detection is improved. Furthermore, the approach can provide an impartial performance comparison of different peak picking algorithms. We also discuss the applicability of the approach to liquid chromatography-mass spectrometry data. References [1] Gloaguen, Y.; Borgsmüller, N. et al. WiPP: Workflow for Improved Peak Picking for Gas Chromatography-Mass Spectrometry (GC-MS) Data. Metabolites 2019, 9, 171.

International audience; We describe herein a Pd-catalyzed methodology for the thioglycoconjugation of iodoaryl peptides and aminoacids. This operationally simple process occurs under semi-aqueous conditions and displays wide substrate scope. The strategy has been successfully applied to both the thioglycosylation of unprotected peptides and the generation of thioglyco-aminoacid building blocks, including those suitable for solid phase peptide synthesis. To demonstrate the broad potential of this technique for late stage functionalization, we successfully incorporated challenging unprotected β-S-GlcNAc- and α-S-GalNAc- derivatives into very long unprotected peptides. This study opens the way to new applications in chemical biology, considering the well-recognized advantages of S-glycosides over O-glycosides in terms of resistance towards both enzymatic and chemical degradation.

The pregnane X receptor (PXR) is the main nuclear receptor regulating the expression of xenobiotic-metabolizing enzymes and is highly expressed in the liver and intestine. Recent studies have highlighted its additional role in lipid homeostasis, however, the mechanisms of these regulations are not fully elucidated. We investigated the transcriptomic signature of PXR activation in the liver of adult wild-type vs. Pxr-/- C57Bl6/J male mice treated with the rodent specific ligand pregnenolone 16&alpha, carbonitrile (PCN). PXR activation increased liver triglyceride accumulation and significantly regulated the expression of 1215 genes, mostly xenobiotic-metabolizing enzymes. Among the down-regulated genes, we identified a strong peroxisome proliferator-activated receptor &alpha, (PPAR&alpha, ) signature. Comparison of this signature with a list of fasting-induced PPAR&alpha, target genes confirmed that PXR activation decreased the expression of more than 25 PPAR&alpha, target genes, among which was the hepatokine fibroblast growth factor 21 (Fgf21). PXR activation abolished plasmatic levels of FGF21. We provide a comprehensive signature of PXR activation in the liver and identify new PXR target genes that might be involved in the steatogenic effect of PXR. Moreover, we show that PXR activation down-regulates hepatic PPAR&alpha, activity and FGF21 circulation, which could participate in the pleiotropic role of PXR in energy homeostasis.

IF 3.188 (2017); International audience; Diet-induced obesity (DIO) is associated with a decreased oral fat detection in rodents. This alteration has been explained by an impairment of the lipid-mediated signaling in taste bud cells (TBC). However, factors responsible for this defect remain elusive. Diet rich in saturated fatty acids is known to elicit a metabolic inflammation by promoting intestinal permeation to lipopolysaccharides (LPS), Gram-negative bacteria-derived endotoxins. To determine whether a local inflammation of the gustatory tissue might explain the obese-induced impairment of the oro-sensory detection of lipids, mice were subjected to a DIO protocol. Using a combination of behavioral tests, transcriptomic analyses of gustatory papillae and biochemical assays, we have found that i) DIO elicits a pro-inflammatory genic profile in the circum-vallate papillae (CVP), known to house the highest density of lingual taste buds, ii) NFkB, a key player of inflammatory process, might play a role in this transcriptomic pattern, iii) plasma LPS levels are negatively correlated with the preference for oily solution, and iv) a chronic infusion of LPS at a level similar to that found in DIO mice is not sufficient to alter the spontaneous preference for fat in lean mice. Taken together these data bring the demonstration that a saturated high fat diet elicits an inflammatory response at the level of peripheral gustatory pathway and a LPS-induced low-grade endotoxemia alone does not explain the change in the preference for dietary lipids observed in DIO mice.

Wrist osteoarthritis (OA) is one of the most common conditions encountered by hand surgeons with limited efficacy of non-surgical treatments. The purpose of this study is to describe the Platelet-Rich Plasma (PRP) mixed-microfat biological characteristics of an experimental Advanced Therapy Medicinal Product (ATMP) needed for clinical trial authorization and describe the clinical results obtained from our first three patients 12 months after treatment (NCT03164122). Biological characterization of microfat, PRP and mixture were analysed in vitro according to validated methods. Patients with stage four OA according to the Kellgren Lawrence classification, with failure to conservative treatment and a persistent daily painful condition &gt, 40 mm according to the visual analog scale (VAS) were treated. Microfat-PRP ATMP is a product with high platelet purity, conserved viability of stromal vascular fraction cells, chondrogenic differentiation capacity in vitro and high secretion of IL-1Ra anti-inflammatory cytokine. For patients, the only side effect was pain at the adipose tissue harvesting sites. Potential efficacy was observed with a pain decrease of over 50% (per VAS score) and the achievement of minimal clinically important differences for DASH and PRWE functional scores at one year in all three patients. Microfat-PRP ATMP presented a good safety profile after an injection in wrist OA. Efficacy trials are necessary to assess whether this innovative strategy could delay the necessity to perform non-conservative surgery.

This article has an erratum: https://doi.org/10.1051/e3sconf/20198802003Présentation d'une communication à la conférence internationale i-DUST 2018 – Inter-Disciplinary Underground Science & Technology (2018/06/04-05), Avignon (FRA); International audience; Groundwater is a major source of water for irrigation of vegetables, especially in the Mediterranean basin. Contamination of aquifer by pathogens has been responsible for numerous disease outbreaks worldwide. Several studies reported that groundwater dissolved organic matter (DOM) can serve as a source of carbon and energy for heterotrophic metabolism of pathogens. In this study, we aimed to investigate the DOM composition of groundwater collected at Avignon. Six liters of groundwater were filtered (0.2 µm) and freeze-dried following appropriate cleaning procedure. The bulk analyses of powder sample were performed using 1D and 2D nuclear magnetic resonance spectroscopy and liquid chromatography coupled with mass spectroscopy. Several components were found at concentrations around 1 µM and comprise : (i) humic and fulvic acids originated from land-derived material or soils and, (ii) various acids, esters and alcohols of different sizes including acetate, lactate and formate, these may result from microbial metabolism. In conclusion, the Avignon groundwater DOM contains a heterogeneous mixture of dissolved organic components with a rather low potential bioreactivity based on the low level of labile biogeochemicals such as carbohydrates.

Biogas plants for waste treatment valorization are presently experiencing rapid development, especially in the agricultural sector, where large amounts of digestate are being generated. In this study, we investigated the effect of vibro-ball milling (VBM) for 5 and 30 min at a frequency of 20 s&minus, 1) on the physicochemical composition and enzymatic hydrolysis (30 U g&minus, 1 total solids (TS) of cellulase and endo-1,4-xylanase from Trichoderma longibrachiatum) of dry and wet solid digestates from an agricultural biogas plant. We found that VBM of dry solid digestate improved the physical parameters as both the particle size and the crystallinity index (from 27% to 75%) were reduced. By contrast, VBM of wet solid digestate had a minimal effect on the physicochemical parameters. The best results in terms of cellulose and hemicelluloses hydrolysis were noted for 30 min of VBM of dry solid digestate, with hydrolysis yields of 64% and 85% for hemicelluloses and cellulose, respectively. For the condition of 30 min of VBM, bioethanol and methane production on the dry solid separated digestate was investigated. Bioethanol fermentation by simultaneous saccharification and fermentation resulted in an ethanol yield of 98 geth kg&minus, 1 TS (corresponding to 90% of the theoretical value) versus 19 geth kg&minus, 1 TS for raw solid digestate. Finally, in terms of methane potential, VBM for 30 min lead to an increase of the methane potential of 31% compared to untreated solid digestate.

Today, chitosan (CS) is probably considered as a biofunctional polysaccharide with the most notable growth and potential for applications in various fields. The progress in chitin chemistry and the need to replace additives and non-natural polymers with functional natural-based polymers have opened many new opportunities for CS and its derivatives. Thanks to the specific reactive groups of CS and easy chemical modifications, a wide range of physico-chemical and biological properties can be obtained from this ubiquitous polysaccharide that is composed of β-(1,4)-2-acetamido-2-deoxy-d-glucose repeating units. This review is presented to share insights into multiple native/modified CSs and chitooligosaccharides (COS) associated with their functional properties. An overview will be given on bioadhesive applications, antimicrobial activities, adsorption, and chelation in the wine industry, as well as developments in medical fields or biodegradability.

Genetically-controlled plant resistance can reduce the damage caused by pathogens. However, pathogens have the ability to evolve and overcome such resistance. This often occurs quickly after resistance is deployed, resulting in significant crop losses and a continuing need to develop new resistant cultivars. To tackle this issue, several strategies have been proposed to constrain the evolution of pathogen populations and thus increase genetic resistance durability. These strategies mainly rely on varying different combinations of resistance sources across time (crop rotations) and space. The spatial scale of deployment can vary from multiple resistance sources occurring in a single cultivar (pyramiding), in different cultivars within the same field (cultivar mixtures) or in different fields (mosaics). However, experimental comparison of the efficiency (i.e. ability to reduce disease impact) and durability (i.e. ability to limit pathogen evolution and delay resistance breakdown) of landscape-scale deployment strategies presents major logistical challenges. Therefore, we developed a spatially explicit stochastic model able to assess the epidemiological and evolutionary outcomes of the four major deployment options described above, including both qualitative resistance (i.e. major genes) and quantitative resistance traits against several components of pathogen aggressiveness: infection rate, latent period duration, propagule production rate, and infectious period duration. This model, implemented in the R package landsepi, provides a new and useful tool to assess the performance of a wide range of deployment options, and helps investigate the effect of landscape, epidemiological and evolutionary parameters. This article describes the model and its parameterisation for rust diseases of cereal crops, caused by fungi of the genus Puccinia. To illustrate the model, we use it to assess the epidemiological and evolutionary potential of the combination of a major gene and different traits of quantitative resistance. The comparison of the four major deployment strategies described above will be the objective of future studies., Author summary There are many recent examples which demonstrate the evolutionary potential of plant pathogens to overcome the resistances deployed in agricultural landscapes to protect our crops. Increasingly, it is recognised that how resistance is deployed spatially and temporally can impact on rates of pathogen evolution and resistance breakdown. Such deployment strategies are mainly based on the combination of several sources of resistance at different spatiotemporal scales. However, comparison of these strategies in a predictive sense is not an easy task, owing to the logistical difficulties associated with experiments involving the spread of a pathogen at large spatio-temporal scales. Moreover, both the durability of a strategy and the epidemiological protection it provides to crops must be assessed since these evaluation criteria are not necessarily correlated. Surprisingly, no current simulation model allows a thorough comparison of the different options. Here we describe a spatio-temporal model able to simulate a wide range of deployment strategies and resistance sources. This model, implemented in the R package landsepi, facilitates assessment of both epidemiological and evolutionary outcomes across simulated scenarios. In this work, the model is used to investigate the combination of different sources of resistance against fungal diseases such as rusts of cereal crops.

International audience; Peripheral neuropathies are subdivided into acquired and hereditary transmitted disorders.Among hereditary peripheral neuropathies, the most frequent is Charcot-Marie-Tooth disease (CMT).We will describe below, in detail, this disorder and its different forms.Charcot-Marie-Tooth disease constitutes a clinically and genetically heterogeneous group ofhereditary motor and sensory peripheral neuropathies. On the basis of electrophysiologic propertiesand histopathology, CMT has been divided into primary peripheral demyelinating (type 1) andprimary peripheral axonal (type 2) neuropathies. The demyelinating neuropathies classified as CMTtype 1, also known as HMSN I, are characterized by severely reduced motor nerve conductionvelocities (NCV) (less than 38 m/s) and segmental demyelination and remyelination with onionbulb formations on nerve biopsy. The axonal neuropathies classified as CMT type 2, also known asHMSN II, are characterized by normal or mildly reduced NCVs and chronic axonal degeneration andregeneration on nerve biopsy. Among the CMT1 group, there are X-linked, autosomal dominant andautosomal recessive forms of CMT.The typical presenting symptom is a weakness of the feet and ankles. The initial physical findingsare depressed or absent tendon reflexes with a weakness of foot dorsiflexion at the ankle. The typicalaffected adult has a bilateral foot drop, symmetrical atrophy of muscles below the knee (stork legappearance), pes cavus, atrophy of intrinsic hand muscles, especially the thenar muscles of the thumb,and absent tendon reflexes in both upper and lower extremities. The life span is not decreased

International audience; The Formyl Peptide Receptor 2 (FPR2) is a novel promising target for the treatment of influenza. During viral infection, FPR2 is activated by annexinA1, which is present in the envelope of influenza viruses; this activation promotes virus replication. Here, we investigated whether blockage of FPR2 would affect the genome trafficking of influenza virus. We found that, upon infection and cell treatment with the specific FPR2 antagonist WRW4 or the anti-FPR2 monoclonal antibody, FN-1D6-AI, influenza viruses were blocked into endosomes. This effect was independent on the strain and was observed for H1N1 and H3N2 viruses. In addition, blocking FPR2signaling in alveolar lung A549 epithelial cells with the monoclonal anti-FPR2 antibody significantly inhibited virus replication. Altogether, these results show that FPR2signaling interferes with the endosomal trafficking of influenza viruses and provides, for the first time, the proof of concept that monoclonal antibodies directed against FPR2 inhibit virus replication. Antibodies-based therapeutics have emerged as attractive reagents in infectious diseases. Thus, this study suggests that the use of anti-FPR2 antibodies against influenza hold great promise for the future.

International audience; Folates, also known as B9 vitamins, serve as donors and acceptors in one-carbon (C1) transfer reactions. The latter are involved in synthesis of many important biomolecules, such as amino acids, nucleic acids and vitamin B5. Folates also play a central role in the methyl cycle that provides one-carbon groups for methylation reactions. The important functions fulfilled by folates make them essential in all living organisms. Plants, being able to synthesize folates de novo, serve as an excellent dietary source of folates for animals that lack the respective biosynthetic pathway. Unfortunately, the most important staple crops such as rice, potato and maize are rather poor sources of folates. Insufficient folate consumption is known to cause severe developmental disorders in humans. Two approaches are employed to fight folate deficiency: pharmacological supplementation in the form of folate pills and biofortification of staple crops. As the former approach is considered rather costly for the major part of the world population, biofortification of staple crops is viewed as a decent alternative in the struggle against folate deficiency. Therefore, strategies, challenges and recent progress of folate enhancement in plants will be addressed in this review. Apart from the ever-growing need for the enhancement of nutritional quality of crops, the world population faces climate change catastrophes or environmental stresses, such as elevated temperatures, drought, salinity that severely affect growth and productivity of crops. Due to immense diversity of their biochemical functions, folates take part in virtually every aspect of plant physiology. Any disturbance to the plant folate metabolism leads to severe growth inhibition and, as a consequence, to a lower productivity. Whereas today's knowledge of folate biochemistry can be considered very profound, evidence on the physiological roles of folates in plants only starts to emerge. In the current review we will discuss the implication of folates in various aspects of plant physiology and development.

International audience; We previously reported that, in multiple sclerosis (MS) patients with a progressive form of the disease, spinal cord periplaques extend distance away from plaque borders and are characterized by the co-occurrence of partial demyelination, astrocytosis and low-grade inflammation. However, transcriptomic analyses did not allow providing a comprehensive view of molecular events in astrocytes vs. oligodendrocytes. Here, we re-assessed our transcriptomic data and performed co-expression analyses to characterize astrocyte vs. oligodendrocyte molecular signatures in periplaques. We identified an astrocytosis-related co-expression module whose central hub was the astrocyte gene Cx43/GJA1 (connexin-43, also named gap junction protein α-1). Such a module comprised GFAP (glial fibrillary acidic protein) and a unique set of transcripts forming a TGFB/SMAD1/SMAD2 (transforming growth factor β/SMAD family member 1/SMAD family member 2) genomic signature. Partial demyelination was characterized by a co-expression network whose central hub was the oligodendrocyte gene NDRG1 (N-myc downstream regulated 1), a gene previously shown to be specifically silenced in the normal-appearing white matter (NAWM) of MS patients. Surprisingly, besides myelin genes, the NDRG1 co-expression module comprised a highly significant number of translation/elongation-related genes. To identify a putative cause of NDRG1 downregulation in periplaques, we then sought to identify the cytokine/chemokine genes whose mRNA levels inversely correlated with those of NDRG1. Following this approach, we found five candidate immune-related genes whose upregulation associated with NDRG1 downregulation: TGFB1(transforming growth factor β 1), PDGFC (platelet derived growth factor C), IL17D (interleukin 17D), IL33 (interleukin 33), and IL12A (interleukin 12A). From these results, we propose that, in the spinal cord periplaques of progressive MS patients, TGFB1 may limit acute inflammation but concurrently induce astrocytosis and an alteration of the translation/elongation of myelin genes in oligodendrocytes.

The human gut microbiota utilizes complex carbohydrates as major nutrients. The requirement for efficient glycan degrading systems exerts a major selection pressure on this microbial community. Thus, we propose that this microbial ecosystem represents a substantial resource for discovering novel carbohydrate active enzymes. To test this hypothesis we screened the potential enzymatic functions of hypothetical proteins encoded by genes of Bacteroides thetaiotaomicron that were up-regulated by arabinogalactan proteins or AGPs. Although AGPs are ubiquitous in plants, there is a paucity of information on their detailed structure, the function of these glycans in planta, and the mechanisms by which they are depolymerized in microbial ecosystems. Here we have discovered a new polysaccharide lyase family that is specific for the L-rhamnose-alpha 1,4-D-glucuronic acid linkage that caps the side chains of complex AGPs. The reaction product generated by the lyase, Delta 4,5-unsaturated uronic acid, is removed from AGP by a glycoside hydrolase located in family GH105, producing the final product 4-deoxy-beta-L-threo-hex-4-enepyranosyl- uronic acid. The crystal structure of a member of the novel lyase family revealed a catalytic domain that displays an (alpha/alpha)(6) barrel-fold. In the center of the barrel is a deep pocket, which, based on mutagenesis data and amino acid conservation, comprises the active site of the lyase. A tyrosine is the proposed catalytic base in the beta-elimination reaction. This study illustrates how highly complex glycans can be used as a scaffold to discover new enzyme families within microbial ecosystems where carbohydrate metabolism is a major evolutionary driver.

International audience; Vitamin E (VE) is the generic term for four tocopherols and four tocotrienols that exhibit the biological activity of alpha-tocopherol. VE status, which is usually estimated by measuring fasting blood VE concentration, is affected by numerous factors, such as dietary VE intake, VE absorption efficiency, and VE catabolism. Several of these factors are in turn modulated by genetic variations in genes encoding proteins involved in these factors. To identify these genetic variations, two strategies have been used: genome-wide association studies and candidate gene association studies. Each of these strategies has its advantages and its drawbacks, nevertheless they have allowed us to identify a list of single nucleotide polymorphisms associated with fasting blood VE concentration and alpha-tocopherol bioavailability. However, much work remains to be done to identify, and to replicate in different populations, all the single nucleotide polymorphisms involved, to assess the possible involvement of other kind of genetic variations, e.g., copy number variants and epigenetic modifications, in order to establish a reliable list of genetic variations that will allow us to predict the VE status of an individual by knowing their genotype in these genetic variations. Yet, the potential usefulness of this area of research is exciting with regard to personalized nutrition and for future clinical trials dedicated to assessing the biological effects of the various isoforms of VE.

Genes acquired by horizontal transfer are increasingly being found in animal genomes. Understanding their origin and evolution requires knowledge about the phylogenetic relationships from both source and recipient organisms. We used RNASeq data and respective assembled transcript libraries to trace the evolutionary history of polygalacturonase (pectinase) genes in stick insects (Phasmatodea). By mapping the distribution of pectinase genes on a Polyneoptera phylogeny, we identified the transfer of pectinase genes from known phasmatodean gut microbes into the genome of an early euphasmatodean ancestor that took place between 60 and 100 million years ago. This transfer preceded the rapid diversification of the suborder, enabling symbiont-free pectinase production that would increase the insects’ digestive efficiency and reduce dependence on microbes. Bacteria-to-insect gene transfer was thought to be uncommon, however the increasing availability of large-scale genomic data may change this prevailing notion.

Two methods for the preparation of 5-iodo-1,2,3-triazolyl-4-phosphonate were explored. This compound was then functionalized by Suzuki and Stille cross-coupling reaction to obtain 5-aryl-, 5-heteroaryl- or 5-alkenyl-1,2,3-triazolyl-4-phosphonates.

The present study aimed at developing an original pre-column HPLC assay allowing rapid characterization of nitric oxide (NO) scavengers from complex plant extracts. Sodium nitroprusside (SNP) was employed as a NO donor and spiked with an aqueous extract from Aloysia triphylla leaves prior to HPLC analysis. Relying on the ability of radical scavenging constituents to be oxidized upon reaction with radicals, this assay successfully allowed direct identification of three potential NO scavengers, including verbascoside, isoverbascoside, and luteolin-7-O-diglucuronide. These three phenolics were also individually assessed for their NO scavenging activities by using a Griess colorimetric assay. With respective IC50 values of 56 &plusmn, 4, 51 &plusmn, 3, and 69 &plusmn, 5 &micro, g/mL, verbascoside, isoverbascoside, and luteolin-7-O-diglucuronide were all reported as potent NO scavenging compounds, confirming the efficiency of the SNP spiking HPLC assay. The present method can, thus, be considered as a valuable and effective approach for speeding up the discovery of NO scavenging constituents.

In mammals, detection of osmolarity by the gustatory system was overlooked until recently. In insects, specific taste receptor neurons detect hypoosmotic stimuli and are commonly called "W" (water) cells. W cells are easy to access in vivo and represent a good model to study the transduction of hypoosmotic stimuli. Using pharmacological and genetic approaches in Drosophila, we show that tarsal W cell firing activity depends on the concentration of external calcium bathing the dendrite. This dependence was confirmed by the strong inhibition of W cell responses to hypoosmotic stimuli by lanthanum (IC(50) = 8 nM), an ion known to inhibit calcium-permeable channels. Downstream, the transduction pathway likely involves calmodulin because calmodulin antagonists such as W-7 (IC(50) = 2 microM) and fluphenazine (IC(50) = 30 microM) prevented the activation of the W cell by hypoosmotic stimuli. A protein kinase C (PKC) may also be involved as W cell responses were blocked by PKC inhibitors, chelerythrine (IC(50) = 20 microM) and staurosporine (IC(50) = 30 microM). It was also reduced when expressing an inhibitory pseudosubstrate of PKC in gustatory receptor neurons. In the rat, the transduction pathway underlying low osmolarity detection involves aquaporin and swelling-activated ion channels. Our study suggests that the transduction pathway of hypoosmotic stimuli in insects differs from mammals.

International audience; This study falls within the framework of the industrial exploitation of by-products of the prickly pear (Opuntia ficus-indica). The study aims to evaluate the use of hydro-ethanolic extract of prickly pear peels as a substitute of vitamin E used as antioxidant in margarine preservation. The extract was rich in total phenolics (1512.58 mg GAE/100 g DM). HPLC-DAD-ESI-MSn analyses allowed the identification of sixteen compounds belonging to hydroxybenzoic acids, hydroxycinnamic acids and flavonoids. The extract displayed a reducing power and an antiradical activity that were respectively similar to and lower than the two antioxidant standards quercetin and butylated hydroxyanisole. Tests conducted at laboratory and pilot scales showed that the margarines elaborated with peel extract were more resistant to oxidation than the margarine reference with vitamin E. In addition, neither the physicochemical nor the microbiological properties were modified. Prickly pear peels contain bioactive substances that could be used in different food sectors.

AGAP : équipe ID; International audience; NMR spectroscopy allows measurements of very accurate values of equilibrium dissociation constants using chemical shift perturbation methods, provided that the concentrations of the binding partners are known with high precision and accuracy. The accuracy and precision of these experiments are improved if performed using individual capillary tubes, a method enabling full automation of the measurement. We provide here a protocol to set up and perform these experiments as well as a robust method to measure peptide concentrations using tryptophan as an internal standard.

International audience; Carrageenans are sulfated polysaccharides extracted from the cell wall of some marine red algae. These polysaccharides are widely used as gelling, stabilizing, and viscosifying agents in the food and pharmaceutical industries. Since the rheological properties of these polysaccharides depend on their sulfate content, we screened several isolated marine bacteria for carrageenan specific sulfatase activity, in the aim of developing enzymatic bioconversion of carrageenans. As a result of the screening, an iota-carrageenan sulfatase was detected in the cell-free lysate of the marine bacterium Pseudoalteromonas carrageenovora strain Psc(T). It was purified through Phenyl Sepharose and Diethylaminoethyl Sepharose chromatography. The pure enzyme, Psc t-CgsA, was characterized. It had a molecular weight of 115.9 kDaltons and exhibited an optimal activity/stability at pH similar to 8.3 and at 40 +/- 5 degrees C. It was inactivated by phenylmethylsulfonyl fluoride but not by ethylene diamine tetraacetic acid. Psc t.-CgsA specifically catalyzes the hydrolysis of the 4-S sulfate of iota-carrageenan. The purified enzyme could transform iota-carrageenan into hybrid iota-/alpha- or pure alpha-carrageenan under controlled conditions. The gene encoding Psc t-CgsA, a protein of 1038 amino acids, was cloned into Escherichia colt, and the sequence analysis revealed that Psc t.-CgsA has more than 90% sequence identity with a putative uncharacterized protein Q3IKL4 from the marine strain Pseudoalteromonas haloplanktis TAC 125, but besides this did not share any homology to characterized sulfatases. Phylogenetic studies show that P carrageenovora sulfatase thus represents the first characterized member of a new sulfatase family, with a C-terminal domain having strong similarity with the superfamily of amidohydrolases, highlighting the still unexplored diversity of marine polysaccharide modifying enzymes.

Background: Peroxisomal ABC transporters are predicted to function as homodimers in mammals. [br/] Results: ABCD1 interacts with ABCD2. Chimeric proteins mimicking full-length dimers represent novel tools for functional study. Artificial homodimers and heterodimers are functional. [br/] Conclusion: Interchangeability between ABCD1 and ABCD2 is confirmed, but PUFA transport depends on ABCD2. [br/] Significance: For the first time, heterodimers in mammals are proven to be functional.[br/] ABCD1 and ABCD2 are two closely related ATP-binding cassette half-transporters predicted to homodimerize and form peroxisomal importers for fatty acyl-CoAs. Available evidence has shown that ABCD1 and ABCD2 display a distinct but overlapping substrate specificity, although much remains to be learned in this respect as well as in their capability to form functional heterodimers. Using a cell model expressing an ABCD2-EGFP fusion protein, we first demonstrated by proximity ligation assay and co-immunoprecipitation assay that ABCD1 interacts with ABCD2. Next, we tested in the pxa1/pxa2 yeast mutant the functionality of ABCD1/ABCD2 dimers by expressing chimeric proteins mimicking homo- or heterodimers. For further structure-function analysis of ABCD1/ABCD2 dimers, we expressed chimeric dimers fused to enhanced GFP in human skin fibroblasts of X-linked adrenoleukodystrophy patients. These cells are devoid of ABCD1 and accumulate very long-chain fatty acids (C26:0 and C26:1). We checked that the chimeric proteins were correctly expressed and targeted to the peroxisomes. Very long-chain fatty acid levels were partially restored in transfected X-linked adrenoleukodystrophy fibroblasts regardless of the chimeric construct used, thus demonstrating functionality of both homo- and heterodimers. Interestingly, the level of C24:6 n-3, the immediate precursor of docosahexaenoic acid, was decreased in cells expressing chimeric proteins containing at least one ABCD2 moiety. Our data demonstrate for the first time that both homo- and heterodimers of ABCD1 and ABCD2 are functionally active. Interestingly, the role of ABCD2 (in homo- and heterodimeric forms) in the metabolism of polyunsaturated fatty acids is clearly evidenced, and the chimeric dimers provide a novel tool to study substrate specificity of peroxisomal ATP-binding cassette transporters.

Microparticulated whey proteins (MWP) in suspension or included in a globular protein network revealed that the degree of flocculation was important in the final structure of the material. The MWP dispersions developed an antithixotropic character when low shear rate was applied to the system even at low temperature. Similar results were obtained on globular protein solutions and could be interpreted in terms of an orthokinetic aggregation. MWP participated to the formation and reinforcement of heat-induced β -lactoglobulin (BLG) gels. The mechanical properties of mixed gels demonstrated synergestic effects at low MWP massic fraction (BLG:MWP ratio higher than 2:1). A possible interpretation would be that both interaction of MWP with the network and flocculation of the MWP inside the pores could occur. However, increasing the fraction of MWP induced heterogeneities in the protein network. In particular, micro-phase separation due to steric hindrance effects or thermodynamic incompatibility between the two components could occur and decrease viscoelastic properties of mixed gels.

Les odeurs résultent la plupart du temps de la perception de mélanges de molécules odorantes. De nombreuses études ont montré que les caractéristiques perceptives des mélanges d’odorants sont souvent différentes de celles de leurs constituants. Ainsi l’intensité odorante d’un mélange peut être supérieure (synergie ou hyper-addition) ou inférieure (hypo-addition) à la simple somme arithmétique des intensités des constituants du mélange. Les mélanges d’odorants peuvent également donner lieu à de nouvelles qualités d’odeurs (perception synthétique) ou laisser apparaître celles des constituants (perception analytique). Cependant, les mécanismes biologiques impliqués dans la perception des mélanges restent méconnus. Quelques études montrent que des interactions à l’entrée du système olfactif pourraient jouer un rôle important dans le traitement de l’information olfactive. En effet, des interactions compétitives et non compétitives ont été observées au niveau des récepteurs olfactifs (RO) et des neurones sensoriels olfactifs (NSO). Dans cette thèse, nous avons étudié en parallèle les réponses des RO et des NSO ainsi que les réponses comportementales à trois mélanges binaires: Octanal (Oct) + Citronnellal (Cit), Oct + Méthional (Méth) et acétate d’isoamyle (ISO) + whisky lactone (WL). Trois approches ont été mises en œuvre: i) des études in vitro des RO par imagerie calcique sur des cellules HEK293; ii) des mesures ex-vivo d’électro-olfactogrammes au niveau des NSO de rat ; iii) des études psychophysiques chez l’Homme. Les résultats ont montré de fortes similitudes entre les réponses des RO/NSO et les réponses perceptives chez l’Homme pour les mélanges Oct + Méth et WL + ISO. Les liens sont moins importants pour le mélange Oct + Cit pour lequel les RO étudiés génèrent des réponses différentes de celles observées aux niveaux plus intégrés. Globalement, ces résultats soutiennent l'hypothèse que les interactions qui se produisent au niveau périphérique contribuent de manière significative au codage olfactif des mélanges odorants, Odours result from the perception of odorant mixtures. Several studies reported that perceptual characteristics of these mixtures are often different from those of their individual components. It has been shown that odour intensity for mixtures can be higher (synergy/hyper-addition) or lower (hypo-addition) than the simple arithmetic sum of each component’s intensity. Besides, mixtures can also give rise to novel odour qualities (configural perception) or conversely are perceived as the juxtaposition of individual components’ odour quality (elemental perception). However, the biological mechanisms that govern odour mixture perception remain unclear, even if there is increasing evidence that peripheral phenomena could play an important role in mixture processing. Indeed, competitive and non-competitive interactions have been observed at the olfactory receptor (OR) and olfactory sensory neuron (OSN) levels. In this PhD work, we investigated in parallel the responses of OR, ON and human behavioural responses for three binary mixtures of odorants: Octanal (Oct) + Citronellal (Cit), Oct + Methional (Meth) and isoamyl acetate (ISO) + whiskey lactone (WL). Three types of experiments were performed: (i) in vitro OR calcium imaging of HEK293 cells; (ii) ex vivo electro-olfactogram measurement on rat OSN; (iii) psychophysical measurements in human. Our results showed clear similarities between the responses of OR/OSN and perceptual responses in human for the two mixtures Oct + Meth and ISO + WL, whereas for the Oct + Cit mixture, OR responses are hardly reconcilable with more integrated responses. Taken together, these findings support the hypothesis that interactions occurring at the peripheral level of the olfactory system contribute significantly to the olfactory coding of odour mixture

Peroxisomes are essential organelles in higher eukaryotes as they play a major role in numerous metabolic pathways and redox homeostasis. Some peroxisomal abnormalities, which are often not compatible with life or normal development, were identified in severe demyelinating and neurodegenerative brain diseases. The metabolic roles of peroxisomes, especially in the brain, are described and human brain peroxisomal disorders resulting from a peroxisome biogenesis or a single peroxisomal enzyme defect are listed. The brain abnormalities encountered in these disorders (demyelination, oxidative stress, inflammation, cell death, neuronal migration, differentiation) are described and their pathogenesis are discussed. Finally, the contribution of peroxisomal dysfunctions to the alterations of brain functions during aging and to the development of Alzheimer's disease is considered.

The objective of this study was to investigate in situ transesterification allowing to produce directly biodiesel from jatropha seed. The influences of amount of KOH catalyst, methanol to seed ratio, amount of n-hexane to methanol and seed ratio, stirring speed, temperature and reaction time were examined to define the best performance of biodiesel yield and quality. Generally, methanol to seed ratio, amount of KOH and n-hexane to methanol and seed ratio affected biodiesel yield. An increase of biodiesel yield was observed as methanol to seed ratio, amount of KOH and n-hexane to methanol and seed ratio were increased. Stirring speed, temperature and reaction time did not affected biodiesel yield. Highest biodiesel yield (89%) was obtained under 6:1 methanol to seed ratio, 0.075 mol/L KOH in methanol, 3:3:1 n-hexane to methanol and seed ratio, 600 rpm stirring speed, 40°C temperature and 6 h reaction time. The effect of process parameters on biodiesel quality was less important. In all experiments tested, the biodiesel quality was very good (acid value < 0.3 mg of KOH/g, viscosity < 5.5 cSt, saponification value > 183 mg of KOH/g). The quality of biodiesel produced under optimal reaction condition was in accordance with the Indonesian Biodiesel Standard.

AtMAP65-4 is a microtubule-associated protein belonging to the AtMAP65s family that comprises 9 members identified in Arabidopsis thaliana. These proteins belong to a family conserved during evolution, MAP65s. Thus, homologous proteins are present in mammals (PRC1), in yeast (Ase1p) or Drosophila (FEO). So far the study of molecular properties and functional AtMAP65s has focused mainly on AtMAP65-1 and AtMAP65-5. The main feature of these proteins is to induce the formation of microtubule bundles in vitro. In vivo, these AtMAP65s are localized with subsets of microtubule bundles as they are suggested to play a role in establishing and maintaining these networks. From the results we obtained on AtMAP65-4 properties during this work such as the in vivo localization, biochemical properties and functional effetc on the MT polymerization, we suggested that the in vivo function of AtMAP65-4 is involved in setting up and maintaining microtubule bundles within kinetochore fibers during cell division. In vitro studies allowed us to show that AtMAP65-4 changes the dynamic parameters of microtubule. In addition to its ability to form bundles, AtMAP65-4 allows steady growth of microtubules in bundles it induces. The mechanism of action of the MAP at the molecular level was analyzed through a bioinformatics study where we modeled the activity of AtMAP65-4 and concluded that it could block the depolymerization events. Moreover, the activity of AtMAP65-4 could be regulated in vivo by post-translational modifications. Indeed, we have shown that AtMAP65-4 is phosphorylated by Aurora kinases in vitro. The effect of this phosphorylation during mitosis is under investigation., AtMAP65-4 est une protéine associée aux microtubules appartenant à la famille des AtMAP65s qui compte 9 membres identifiés chez Arabidopsis thaliana. Ces protéines appartiennent à une famille conservée au cours de l'évolution, les MAP65s. Ainsi, des protéines homologues sont présentes chez de mammifères (PRC1), chez la levure (Ase1p) ou chez la drosophile (FEO). Jusqu'ici l'étude des propriétés moléculaires et fonctionnelles des AtMAP65s s'est portée essentiellement sur l'étude d'AtMAP65-1 et AtMAP65-5. La principale caractéristique de ces protéines est d'induire la formation de faisceaux de microtubules in vitro. La distribution des AtMAP65s in vivo est très régulée, celle-ci sont localisées avec des réseaux des microtubules bien définis. Ainsi, leur rôle supposé est de mettre en place ces réseaux puis de participer à leur maintient. La localisation d'AtMAP65-4 apparait comme très intéressante car elle est strictement associée avec les microtubules du fuseau mitotique. D'après les résultats obtenus au cours de ce travail, nous avons suggéré que la fonction in vivo d'AtMAP65-4 est de participer à la mise en place et au maintient des microtubules en faisceaux dans les fibres kinétochoriennes lors de la division cellulaire. Lors d'une étude in vitro nous avons montré qu'AtMAP65-4 modifie les paramètres dynamiques de polymérisation des microtubules. Outre sa capacité à former des faisceaux, AtMAP65-4 permet une croissance régulière des microtubules au sein des faisceaux qu'elle induit. Le mécanisme d'action de la MAP à l'échelle moléculaire a été analysé à travers une étude bioinformatique où nous avons modélisé l'activité d'AtMAP65-4. Les données obtenues montrent qu'AtMAP65-4 peut bloquer les évènements de dépolymérisation des microtubules. Par ailleurs, l'activité d'AtMAP65-4 pourrait être régulée in vivo par des modifications post traductionnelles. En effet, nous avons montré et étudié l'effet de la phosphorylation d'AtMAP65-4 par les kinases Auroras. Cette phosphorylation pourrait être impliquée dans la régulation de l'activité d'AtMAP65-4 au cours de la mitose.

author can archive publisher's version/PDF (embargo 12 mois); Uptake, retention and release of 5 selected odorants (benzaldehyde, 2-methylpyrazine, 2-isobutyl-3-methoxypyrazine, 2-isobutylthiazole, and 2,4,5-trimethylthiazole) by recombinant rat odor-binding protein 3 (rat-OBP3) were measured in a model system under nonequilibrium conditions. Gaseous odorants were introduced into a 100 mm section of a polar deactivated capillary in which aqueous rat-OBP3 films were formed to mimic the olfactory epithelium (OE), and the change in the gas-phase concentration of the outflow gas was monitored in real time using atmospheric pressure chemical ionization-mass spectrometry (APCI-MS). The 5 odorants were chosen because they exhibited a broad range of dissociation constants with rat-OBP3 and because they were amenable to detection by on-line APCI-MS. All 5 odorants were quantitatively bound by rat-OBP3, which resulted in an effective concentration of the odorants in the aqueous layer (about 50 000-fold). Odorant release from the rat-OBP3-odorant complex into the gas phase showed that odorant release was governed by the dissociation constant of the complex and the flow rate of odorant-free air. When 2 odorants were introduced into the system, odorant uptake and release were influenced by the method of introduction and their relative affinities for the protein. Because rat-OBP3 exhibits typical odorant-binding characteristics, the results not only provide fundamental information on the kinetics of odorant mass transfer induced by the presence of OBPs in the olfactory mucus layer but also support the possibility that vertebrate OBPs may facilitate the accumulation of odorants in the OE.

International audience; The enzymic degradation of plant cell walls plays a central role in the carbon cycle and is of increasing environmental and industrial significance. The catalytic modules of enzymes that catalyze this process are generally appended to noncatalytic carbohydrate-binding modules (CBMs). CBMs potentiate the rate of catalysis by bringing their cognate enzymes into intimate contact with the target substrate. A powerful plant cell wall-degrading system is the Clostridium thermocellum multienzyme complex, termed the "cellulosome." Here, we identify a novel CBM (CtCBM62) within the large C. thermocellum cellulosomal protein Cthe_2193 (defined as CtXyl5A), which establishes a new CBM family. Phylogenetic analysis of CBM62 members indicates that a circular permutation occurred within the family. CtCBM62 binds to d-galactose and l-arabinopyranose in either anomeric configuration. The crystal structures of CtCBM62, in complex with oligosaccharides containing α- and β-galactose residues, show that the ligand-binding site in the β-sandwich protein is located in the loops that connect the two β-sheets. Specificity is conferred through numerous interactions with the axial O4 of the target sugars, a feature that distinguishes galactose and arabinose from the other major sugars located in plant cell walls. CtCBM62 displays tighter affinity for multivalent ligands compared with molecules containing single galactose residues, which is associated with precipitation of these complex carbohydrates. These avidity effects, which confer the targeting of polysaccharides, are mediated by calcium-dependent oligomerization of the CBM.

Les couleurs rencontrées dans les fruits et légumes résultent de l’accumulation de molécules colorées appelées pigments. On en trouve une immense diversité dans le monde végétal. On peut classer les pigments en deux grandes catégories : ceux solubles dans l’eau (anthocyanes et flavonols) et ceux solubles dans l’huile (caroténoïdes et chlorophylles). Certains pigments peuvent être extraits par divers procédés et peuvent ainsi servir de colorant alimentaire, défini par un code E .

International audience; How information is transformed along synaptic processing stages is critically important to understand the neural basis of behavior in any sensory system. In moths, males rely on sex pheromone to find their mating partner. It is essential for a male to recognize the components present in a pheromone blend, their ratio, and the temporal pattern of the signal. To examine pheromone processing mechanisms at different levels of the olfactory pathway, we performed single-cell recordings of olfactory receptor neurons (ORNs) in the antenna and intracellular recordings of central neurons in the macroglomerular complex (MGC) of the antennal lobe of sexually mature Agrotis ipsilon male moths, using the same pheromone stimuli, stimulation protocol, and response analyses. Detailed characteristics of the ORN and MGC-neuron responses were compared to describe the transformation of the neuronal responses that takes place in the MGC. Although the excitatory period of the response is similar in both neuron populations, the addition of an inhibitory phase following the MGC neuron excitatory phase indicates participation of local interneurons (LN), which remodel the ORN input. Moreover, MGC neurons showed a wider tuning and a higher sensitivity to single pheromone components than ORNs.

International audience; To clarify the role of phospholipase C (PLC) in insect olfactory transduction, we have undertaken its molecular identification in the moth Spodoptera littoralis. From the analysis of a male antennal expressed sequence tag library, we succeeded in cloning a full-length cDNA encoding a PLC that belongs to the cluster of PLC-b subtypes. In adult males, the PLC-b transcript was located predominantly in brain and antennae where its presence was detected in the olfactory sensilla trichodea. Moreover, PLC-b was expressed in antennae at the beginning of the pupal stage, then reached a maximum at the end of this stage and was maintained at this level during the adult period. Taken together, these results provided molecular evidence for the putative participation of a PLC-b in signaling pathways responsible for the establishment and the functioning of insect olfactory system.

International audience; This study presents experimental results on the interaction between smectite clays (nontronite and MX80 bentonite) and the facultative anaerobic, heterotrophic Shewanella putrefaciens in two types of conditions: i) batch experiments with low solid to liquid ratios and agitated oxygenated conditions and ii) reaction-cell experiments with high solid to liquid ratios in anaerobic, confined volume conditions. The former was chosen to study the ability of S. putrefaciens to live on smectite as the only substrate and the latter was designed to simulate more compacted subsurface environments of underground repository waste sites. Bacterial cell counts in the batch experiments reveal the prolonged survival of S. putrefaciens in the smectite suspension compared to standard laboratory culture media. In the case of nontronite, variations in solution chemistry indicate bacterial consumption and/or partial binding of cations. Microscopic investigations show associated biofilm-smectite aggregates and Si-rich gels produced by the partial dissolution of clay mineral grains. In contrast, the MX80 bentonite was not seen to be chemically affected by bacterial activity in batch cultures. However, the confined volume experiments, using reaction-cell X-ray diffraction combined with peak calculations (CALCMIX), do indicate that S. putrefaciens has a pronounced effect on the water content of compacted MX80 bentonite. The presence of these bacteria enhances both the amount of adsorbed interlayer water and the available pore space. The anaerobic conditions were also favourable for accessory phase dissolution (notably calcite) and synchronous precipitation of lepidocrocite related to bacterially induced changes in pH and Eh. The varied response of the two studied clays to the presence of bacteria is attributed largely to the materials composition. The interlayer Ca of nontronite facilitates bacterial attachment to surfaces and Fe(III) provokes the production of chelators that enhance mineral dissolution. Although MX80 bentonite is less affected by bacterially enhanced dissolution, it is more sensitive to microstructural changes. Mechanisms involve aggregation of Na-smectite particles in voids created by cell lyses, the initial production of biofilm and the pH and Eh dependent dissolution and precipitation of accessory minerals. This investigation highlights the importance of including bacteria-mineral studies in assessing the safety issue of underground disposal of nuclear waste material.

International audience; In moths, the components of the female pheromone blend are detected in the male antennae by pheromone olfactory receptor neurons (Ph-ORNs) expressing narrowly tuned olfactory receptors. Responses to sex pheromones have generally been thought to be independent from the odorant background. However, interactions between pheromone components and plant volatiles have been reported at behavioral and detection levels. To document the mechanisms of such interactions, we analyzed Ph-ORN responses of Spodoptera littoralis to the main pheromone component, Z9E11-14:Ac, in the presence of 4 monoterpenes. To mimic natural contexts in which plant odors and pheromone emanate from different sources, the 2 stimuli were presented with different temporal patterns and from independent sources. Linalool reversibly reduced the firing response to Z9E11-14:Ac and produced an off effect. Geraniol and geranyl and linalyl acetates reduced the responses to Z9E11-14:Ac with a longer time course. Pulses of linalool over prolonged pheromone stimulation resulted in a discontinuous firing activity. Pulses of pheromone were better separated over a background of linalool, compared with odorless air. The data confirm that plant compounds may modulate the intensity and the temporal coding by Ph-ORNs of pheromone information. This modulation might positively affect mate location at high pheromone density especially nearby a pheromone source.

In flavor perception, both experience with the components of odor/taste mixtures and the cognitive strategy used to examine the interactions between the components influence the overall mixture perception. However, the effect of these factors on odor mixtures perception has never been studied. The present study aimed at evaluating whether 1) previous exposure to the odorants included in a mixture or 2) the synthetic or analytic strategy engaged during odorants mixture evaluation determines odor representation. Blending mixtures, in which subjects perceived a unique quality distinct from those of components, were chosen in order to induce a priori synthetic perception. In the first part, we checked whether the chosen mixtures presented blending properties for our subjects. In the second part, 3 groups of participants were either exposed to the odorants contributing to blending mixtures with a "pineapple" or a "red cordial" odor or nonexposed. In a following task, half of each group was assigned to a synthetic or an analytical task. The synthetic task consisted of rating how typical (i.e., representative) of the target odor name (pineapple or red cordial) were the mixtures and each of their components. The analytical task consisted of evaluating these stimuli on several scales labeled with the target odor name and odor descriptors of the components. Previous exposure to mixture components was found to decrease mixture typicality but only for the pineapple blending mixture. Likewise, subjects engaged in an analytical task rated both blending mixtures as less typical than did subjects engaged in a synthetic task. This study supports a conclusion that odor mixtures can be perceived either analytically or synthetically according to the cognitive strategy engaged.

Perceptual interactions in a model of wine woody–fruity binary mixtures were previously reported in a psychophysical study performed through orthonasal stimulation only. However, recent studies suggested that the perception of food-like and nonfood-like odors may depend on the route of stimulation. The aim of the present study was two-fold: first to examine the neural correlates of perceptual interactions using electroencephalogram (EEG)-derived event-related potentials (ERPs) and second to test the influence of the stimulation route on quality perception. Therefore, we designed an experiment with 30 subjects to study perceptual interactions in woody–fruity mixtures and compared ortho- vs. retronasal stimulation sites on perceived odor quality and ERPs. The results revealed synergy or masking of the fruity component, depending on the woody component level. Synergy was supported by larger N1 amplitude of the ERP. Furthermore, mixtures including a medium level of the woody odor elicited a strong increase of P2 amplitude only retronasally. This study evidenced for the first time electrophysiological correlates of both perceptual synergy and masking on the early component of the ERPs and confirmed that retro- vs. orthonasal stimulation route induces different neural processes that are reflected in the late component of the ERP.

International audience; The judgment of pleasantness/unpleasantness is the prominent reaction to the olfactory world. In human adults, the hedonic valence of odor perception is affected by various factors, among which is an individual's lexical knowledge about smells. The present study examined whether such top-down effects of lexical knowledge on hedonic judgment of olfactory input are similar in children (5-6 years) and adults (20-25 years). In both groups, the lexical knowledge was found to influence the perception of the least emotional (or most neutral) odors: the pleasantness of the smells of banana and mint was enhanced when participants were given the corresponding odor label before olfactory sensation. These results lend support to the notion that, during childhood, smells are not only encoded perceptually but that verbal encoding also steers contextual effects that may be prominent factors in the early memorization and categorization of odors.

International audience; We asked whether the large variability in odor imaging ability is underlain by interindividual differences in the processing of smells and emotion. Olfactory imaging ability, anhedonia level, and odor perception were measured in 40 subjects, using the Vividness of Olfactory Imagery Questionnaire (VOIQ), the Physical Anhedonia Scale, and the European Test of Olfactory Capabilities. "Good" olfactory imagers, defined primarily on the basis of the VOIQ, rated pleasant smells as more familiar and had lower anhedonia scores than "bad" olfactory imagers. Based on self-reported measures, these results suggest that, like olfactory perception, the mental imagery of smells is related to emotion and that, beyond their differences in vividness, good and bad olfactory imagers differ in their experience of emotion and long-term memory of smells.

N° 261; The role of hedonic reactions to odours on determining rejections of foods in infants is still debated. To answer this question, the first requirement is to have an instrument to test infants’ olfactory reactivity. Indeed, except in neonates, there is a lack of validated method. This study presents results of a new method designed to assess olfactory reactivity of 7- to 8-month-old infants. This method is based on the observation of infants’ reactions while they explored different rattles. The rattles were visually identical but differed in their odours (neutral or with a food odour). Two sequences of 6 rattles were presented to each infant. Eight odorants were used. Four odorants were chosen to evoke odours of often rejected foods and four were chosen to evoke odours of often liked foods. The hedonic valence was previously checked with an adult panel. These odours were also equated for intensity and had a very low perceived trigeminal component. Forty infants were tested and videotaped. These videotapes were subjected to frame-by-frame analysis to measure a variety of behaviours supposed to reflect the infants’ hedonic reaction (e.g., times spent to mouthing or pushing away the rattle). Duration of these events for each odorized rattle was compared to those observed for the control rattles. It thus allows getting a reactivity index per odorant and over all odorants. Reactivity of infants to these odorants was related to infants’ reactions to food during weaning, evaluated by the mother. Methodological considerations concerning the reliability of measures of hedonic reactivity in infants will be discussed. The issue of whether young children experience odours as pleasant or unpleasant will be discussed in relation with their food experiences.

International audience

A peptide nitrile hydratase activity has been engineered into the cysteine protease papain by a single carefully selected mutation at the active site of the enzyme. The papain variant Gln19Glu hydrolyzes the substrate MeOCO-PheAla-CN to the corresponding amide with a kcat/KM value of 1.15 x 10(3) M-1 s-1. The reaction leads to an accumulation of the corresponding amide, which is then further hydrolyzed to the acid by the natural amidase activity of the enzyme. The pH-dependency of the nitrile hydratase activity of Gln19Glu supports the involvement of the acid form of the Glu19 residue in the reaction. The wild type enzyme displays very weak nitrile hydratase activity, and the introduction of a glutamic acid residue in the oxyanion hole of papain causes the kcat at pH 5 to increase by a factor of at least 4 x 10(5). Peptide nitriles react with cysteine proteases to form thioimidates, and the role of the glutamic acid residue introduced at position 19 in the Gln19Glu enzyme is to participate in the acid-catalyzed hydrolysis of the thiomidate to the amide by the provision of a proton to form the more reactive protonated thioimidate. This dramatically decreases the energy barrier for the hydrolysis of the thioimidate, as shown by the impressive increase in kcat. The success of the rational approach undertaken is a consequence of the level of understanding of the basic catalytic properties of cysteine proteases of the papain family.

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International audience; The matricial and part of the fibrillar polysaccharides from the cell walls of Gracilaria crassissima and G. tikvahiae were investigated using a sequential solvent extraction, 13C, !H nuclear magnetic resonance and infrared spectroscopy, anion exchange fractionation, and X-ray diffractometry. The gel strength of agar fractions from (7. crassissima were also measured. Agar from G. crassissima, composed essentially of agarobiose repeating units with its biological precursor, was extracted with cold water and boiling 60% ethanol. The content of 6-O-methylated agarobiose repeating units in this agar was low, and the substituted disaccharide was principally concentrated in one highly methylated fraction extracted from the alga with boiling 80% ethanol. Alkali treatment of fractions converted the mainly charged molecules to neutral agarose that eluted essentially with distilled water from the anionexchanger. The gel strength, 3,6-anhydrogalactose and sulfate contents of alkali-treated fractions were comparable or better than commercial agar or agarose. Thus, G. crassissima represents an excellent potential commercial resource for agar and agarose. In contrast, with G. tikvahiae the 6-0-methylated agar-containing 4-O-methyl L-galactode, was the principal extract in water at 121 °C, probably reflecting cross-linkages of cell-wall polymers. Alkali treatment of this fraction did not affect its ehition pattern through DEAE-Sephadex, demonstrating that the charge on this agar was not associated with biological precursor repeating units. Analysis of the I3C NMR spectra of the polymers revealed resonances that were assigned to the 4-O-methyl α-L-galactopyranosc present in this agar. Anion exchange chromatography of agar fractions from these two algae and the different gel strengths obtained between alkali-treated fractions from G. crassissima indicated that the extracts obtained by the sequential solvent method were heterogeneous. A combination of sequential solvent extraction of agar and anion exchange chromatography is proposed for obtaining better chemically defined agaroses. Finally, X-ray diffractometry definitively demonstrated the presence of cellulose in the fibrous material recovered after extraction of agar from these two algae.